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Method For Producing An Acellular Dermal Matrix, And Acellular Dermal Matrix Produced By Same

a technology of which is applied in the field of preparing acellular dermal matrix and acellular dermal matrix therefr, can solve the problems of increasing patient's pain, skin tissue may be obtained, and regions can leave new scars, so as to achieve the effect of reducing the risk of infection

Active Publication Date: 2012-12-27
CG BIO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a new method for making an acellular dermal matrix that is more stable and has less changes in its biological properties compared to conventional methods.

Problems solved by technology

However, when burnt areas are extensive, there is a limitation of the regions from which skin tissue may be obtained, and the harvesting regions can leave a new scar.
However, harvesting autograft tissue creates a new injury, increasing patient's pain, time for complete recovery can be extended, and the economic burden is greater.
In addition, when insufficient healthy regions remain—as with a severely burned patient—autograft cannot be applied, or grafting operations should be performed repeatedly.
However, other side effects as well as immunorejection often result.
However, tissues prepared according to the method disclosed in the above patent have problems in that cryoprotectant ingredients do not sufficiently penetrate into collagen tissues, and low concentration of sugar cannot sufficiently exclude moisture from collagen tissues—which has a characteristic of absorbing lots of moisture—so that many ice crystals are formed at the time of freezing.

Method used

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  • Method For Producing An Acellular Dermal Matrix, And Acellular Dermal Matrix Produced By Same
  • Method For Producing An Acellular Dermal Matrix, And Acellular Dermal Matrix Produced By Same
  • Method For Producing An Acellular Dermal Matrix, And Acellular Dermal Matrix Produced By Same

Examples

Experimental program
Comparison scheme
Effect test

experimental example 1

Histological Examination

[0066]H&E (hematoxylin & eosin) staining was carried out as follows:

[0067](1) A paraffin block was cut with 4 μm thickness and dried to obtain a paraffin section.

[0068](2) For deparaffinization, after conducting xylene treatment of 5 minutes three times, 100% ethanol treatment of 2 minutes three times, 90% ethanol treatment of 1 minute one time, 80% ethanol treatment of 1 minute one time and 70% ethanol treatment of 1 minute one time, the section was rinsed in running water for 10 minutes.

[0069](3) After staining with hematoxylin for 10 minutes, the section was rinsed in running water for 3 minutes. Then, after staining with eosin for 10 minutes, the section was rinsed in running water until no eosin was detected in the rinse water. After conducting 70% ethanol treatment of 1 second ten times, 80% ethanol treatment of 1 second ten times, 90% ethanol treatment of 1 second ten times, 100% ethanol treatment of 1 minute two times and xylene treatment of 3 minutes...

experimental example 2

Measurement of Degradability by Collagenase

[0080]To evaluate the stability of acellular dermal matrixes of Example and Comparative Examples 1 and 2, the degradability by collagenase was measured as follows:

[0081](1) 25 mg of sample was added to 5 mM TES buffer containing 0.36 mM calcium chloride and mixed well.

[0082](2) 0.1 ml of collagenase (0.1 mg / ml) was added to the sample of step (1) and incubated at 37° C. for one day with stirring.

[0083](3) 4.0 mM L-leucine standard solution was serially diluted and treated with ninhydrin color reagent. A standard curve was prepared by measuring absorbance at 570 nm (VERSA max, Molecular Device, USA).

[0084](4) The sample of step (2) was treated with ninhydrin color reagent and absorbance at 570 nm was then measured.

[0085](5) The amount of released L-leucine from each sample was calculated by using the L-leucine standard curve of step (3).

[0086]The above calculated L-leucine release amount is represented in FIG. 4. As can be seen from FIG. 4, ...

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Abstract

The present invention relates to a method for producing an acellular dermal matrix, and to an acellular dermal matrix produced by same, and more particularly, to a method for producing an acellular dermal matrix, in which sucrose is added to base ingredients consisting of glycerol, propylene glycol, and a base solvent or solution so as to produce a cryoprotectant, the solution is injected into the skin below the epidermis and dermis from which cells have been removed, and freeze-drying is then performed.

Description

TECHNICAL FIELD[0001]The present invention relates to a method for preparing an acellular dermal matrix (ADM) and an acellular dermal matrix prepared therefrom. More specifically, the present invention relates to a method in which a cryoprotectant is prepared by adding sucrose to basic constituents comprising glycerol, propylene glycol, and a basic solvent or solution, and the resulting solution is then penetrated into skin tissue in which epidermis and cells in dermis are removed to prepare an acellular dermal matrix.BACKGROUND ART[0002]Skin is the largest organ, covering the entire human body, and has functions of preventing loss of body fluid, influx of toxic substances and microbes from the outside, and protecting the body from physical and chemical stimuli. In the case of a patient whose skin is seriously impaired by severe burns, injury, carcinoma excision, skin diseases and the like, a protective membrane is needed to prevent infection of impaired regions and the loss of body...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/071
CPCA61L27/362A61L27/3687A61L2430/40A61L27/60
Inventor CHUN, WOOKCHOI, WEON IKHONG, JOON PIORYU, HYUN SEUNG
Owner CG BIO CO LTD
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